| Sirt1基因shRNA干扰载体构建及其对细胞增殖和凋亡的影响 |
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| 引用本文： | 杜文婷,任思冲,所起凤,杨鸣鸣,何丹,刘戟. Sirt1基因shRNA干扰载体构建及其对细胞增殖和凋亡的影响[J]. 生物医学工程学杂志, 2011, 0(5) |
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| 作者姓名： | 杜文婷 任思冲 所起凤 杨鸣鸣 何丹 刘戟 |
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| 作者单位： | 四川大学华西基础医学与法医学院生物化学与分子生物教研室;四川中医药高等专科学校药学与检验系医学检验教研室; |
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| 基金项目： | 教育部博士点基金新教师项目资助(20070610124);教育部留学回国人员基金资助项目(2008890-19-11) |
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| 摘 要： | 本实验目的在于构建Sirt1 shRNA干扰载体,探讨Sirt1对HepG2、A549和293T细胞增殖和凋亡的影响。设计并合成Sirt1的shRNA序列,重组到pGenesil-1.0质粒载体中筛选出pGenesil-1.0-Sirt1阳性克隆并测序。将构建好载体转染HepG2、A549和293T细胞,用RT-PCR、Western blot检测各细胞中Sirt1表达水平,MTT测细胞增殖活性,加入阿霉素处理三组细胞后,用MTT检测细胞凋亡情况。结果表明成功构建Sirt1 shRNA干扰载体,各细胞株中Sirt1表达水平明显下降,Sirt1 shRNA克隆细胞株增殖能力降低,阿霉素处理后,各细胞株抗凋亡能力明显减弱,Sirt1在维持细胞增殖及抗细胞DNA损伤凋亡方面发挥重要作用,为进一步研究提供理论依据。
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| 关 键 词： | 人类Sirt1 shRNA干扰载体 细胞增殖 细胞凋亡 |
Construction of Sirt1 shRNA Interfering Vector and Its Effects on Cell Proliferation and Apoptosis |
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| Du Wenting Ren Sichong Suo Qifeng Yang Mingming He Dan, Liu Ji. Construction of Sirt1 shRNA Interfering Vector and Its Effects on Cell Proliferation and Apoptosis[J]. Journal of biomedical engineering, 2011, 0(5) |
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| Authors: | Du Wenting Ren Sichong Suo Qifeng Yang Mingming He Dan Liu Ji |
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| Affiliation: | Du Wenting1 Ren Sichong1 Suo Qifeng1 Yang Mingming1 He Dan1,2 Liu Ji11(Department of Biochemistry and Molecular Biology,West China School of Preclinical and Forensic Medicine,Sichuan University,Chengdu 610041,China)2(Department of Pharmacy and Medical Laboratory Science,Sichuan College of Traditional Chinese Medicine,Mianyang 621000,China) |
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| Abstract: | This study was aimed to construct Sirt1 shRNA interfering vector and to analyze the effects of Sirt1 on cell proliferation and apoptosis in HepG2,A549 and 293T cell lines.To design and synthisize Sirt1 shRNA sequence then recombinate it to pGenesil-1.0 plasmid,the positive pGenesil-1.0-Sirt1 vector clone was screened by effective detections and sequencing.The vectors were transfected into HepG2,A549,293T cell lines,and Sirt1 expression levels in these clones were detected by RT-PCR and Western-blot.These cl... |
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| Keywords: | Human Sirt1 shRNA interfering vector Cell proliferation Cell apoptosis |
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